The Monomer Covalently Closed Linear Replicative Form DNA of Aleutian Disease Parvovirus is Infectious After Transfection into Permissive Cells

Abstract

Summary The recently described monomer covalently closed linear replicative form DNA (Mccl RF DNA) of Aleutian disease parvovirus (ADV) is an infectious intermediate of the viral DNA replication cycle. Transfection of highly purified Mccl RF DNA into susceptible feline kidney cells (CCC clone 81 cells) resulted in viral DNA replication, expression of viral proteins and synthesis of infectious progeny virus. Mccl RF DNAs generated under permissive (32 °C) or non‐permissive (37 °C) conditions were shown to be biologically indistinguishable. The accumulation of the Mccl RF DNA form at the non‐permissive temperature in vitro strongly resembles that in bone marrow cells of naturally infected mink and may reflect one mechanism contributing to virus persistence of ADV in vivo. Zusammenfassung Die monomer kovalent‐geschlossene lineare replikative Form (Mccl RF)‐DNA des Aleutian Disease Parvovirus (ADV) ist ein infektiöses Intermediat im Rahmen der viralen DNA‐Replikation. Transfektion hochgereinigter Mccl RF DNA in feline Nierenzellen CCC clone 81 induzierte virale DNA‐Replikation, die Expression viraler Proteine und die Synthese infektiöser Virionen. Mccl RF DNAs aus infizierten Zellen inkubiert unter permissiven (32 °C) oder nicht‐permissiven (37 °C) Bedingungen waren funktionell nicht unterscheidbar. Die relative Anhäufung dieser RF‐DNA‐Form unter der nicht‐permissiven Temperatur in vitro ähnelt stark der in Knochenmarkszellen infizierter Nerze in vivo, und ist möglicherweise eine Ursache der Viruspersistenz im Rahmen der Aleutenkrankheit der Nerze.

Specific initiation of replication at the right-end telomere of the closed species of minute virus of mice replicative-form DNA

“…Thus, the cellular extract appears to have little capacity, by itself, to unfold and copy the right-end genomic hairpin which becomes ligated to the growing complementary strand. The cRF species was recently detected in cells infected with the autonomous parvoviruses MVM (13) and Aleutian disease virus (35) and was postulated to be an early intermediate in parvovirus DNA replication (17,56). Our data argue for this possibility by showing that cRF DNA is the direct product of the conversion reaction taking place under in vitro conditions.…”